Team:DTU-Denmark/Notebook/22 August 2013

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===nanodrop measurement of ara spl plasmids===
==Results==
==Results==

Revision as of 16:23, 22 August 2013

22 August 2013

Contents

lab 208


Main purpose


Who was in the lab


Kristian, Henrike

Procedure


USER ligation and transformation

Redid for HAO and cyc in arabinose inducible pZA21::araBAD and for Nir fragments

PCR for Biobrick parts

Set up a new PCR reaction for Biobrick parts using HF buffer and 5% DMSO but no MgCl2. PCR was run on a touchdown program

primers: 53a, 53b

template: Sec2 miniprep

program:

temperature time cycles
98C 2:00 -
98C 0:10 10
63C 1:00 10
-0.5C per cycle
72C 1:00 10
98C 0:10 25
53C 1:00 25
72C 1:00 25
72C 5:00 -
10C hold -

Gel purification

Gel purified the linearized plasmid pZA21::ara with USER endings

colony pPCR to verify AMO insert

Picked 10 colonies from yesterday's cloning for colony PCR. Diluted cells in 50 uL of MilliQ and took 1 uL as template

primers: 53a, 53b

program:

temperature time cycles
98C 10:00 -
98C 0:10 36
56C 0:30 36
72C 2:00 36
72C 5:00 -
10C hold -

nanodrop measurement of ara spl plasmids

Results


Conclusion


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